Sperm from beta1,4-galactosyltransferase I-null mice exhibit precocious capacitation
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Updated: Aug 29, 2026

Using an Extracellular Flux Analyzer to Measure Changes in Glycolysis and Oxidative Phosphorylation during Mouse Sperm Capacitation
Published on: January 22, 2020
Carey Rodeheffer1, Barry D Shur
1Department of Cell Biology, Graduate Program in Biochemistry, Cell and Developmental Biology, Emory University School of Medicine, 615 Michael Street, Atlanta, GA 30322, USA.
Abstract:
Mammalian sperm must undergo a physiological maturation, termed capacitation, before they are able to fertilize eggs. Despite its importance, the molecular mechanisms underlying capacitation are poorly understood. In this paper, we describe the capacitation phenotype of sperm lacking the long isoform of beta1,4-galactosyltransferase I (GalT I), a sperm surface protein that functions as a receptor for the zona pellucida glycoprotein, ZP3, and as an inducer of the acrosome reaction following ZP3-dependent aggregation. As expected, wild-type sperm must undergo capacitation in order to bind the zona pellucida and undergo a Ca(2+) ionophore-induced acrosome reaction. By contrast, GalT I-null sperm behave as though they are precociously capacitated, in that they demonstrate maximal binding to the zona pellucida and greatly increased sensitivity to ionophore-induced acrosome reactions without undergoing capacitation in vitro. The loss of GalT I from sperm results in an inability to bind epididymal glycoconjugates that normally maintain sperm in an 'uncapacitated' state; removing these decapacitating factors from wild-type sperm phenocopies the capacitation behavior of GalT I-null sperm. Interestingly, capacitation of GalT I-null sperm is independent of the presence of albumin, Ca(2+) and HCO(3)(-); three co-factors normally required by wild-type sperm to achieve capacitation. This implies that intracellular targets of albumin, Ca(2+) and/or HCO(3)(-) may be constitutively active in GalT I-null sperm. Consistent with this, GalT I-null sperm have increased levels of cAMP that correlate closely with both the accelerated kinetics and co-factor-independence of GalT I-null sperm capacitation. By contrast, the kinetics of protein tyrosine phosphorylation and sperm motility are unaltered in mutant sperm relative to wild-type. These data suggest that GalT I may function as a negative regulator of capacitation in the sperm head by suppressing intracellular signaling pathways that promote this process.
Insights
Beta1,4-galactosyltransferase I (GalT I) normally inhibits sperm capacitation. Loss of GalT I in sperm leads to precocious capacitation, suggesting GalT I acts as a negative regulator of this crucial fertilization process.
Area of Science:
- Reproductive Biology
- Molecular Biology
- Cell Biology
Background:
- Mammalian sperm require capacitation for egg fertilization, but its molecular mechanisms remain unclear.
- Beta1,4-galactosyltransferase I (GalT I) is a sperm surface protein involved in zona pellucida binding and acrosome reaction induction.
- Understanding GalT I's role is crucial for deciphering sperm maturation pathways.
Purpose of the Study:
- To investigate the role of the long isoform of beta1,4-galactosyltransferase I (GalT I) in mammalian sperm capacitation.
- To characterize the capacitation phenotype of sperm lacking GalT I.
- To elucidate the molecular mechanisms by which GalT I influences sperm maturation.
Main Methods:
- Generation and analysis of GalT I-null sperm.
- Assessment of sperm-zona pellucida binding.
- Evaluation of calcium ionophore-induced acrosome reactions.
- Analysis of epididymal glycoconjugate binding.
- Investigation of capacitation co-factor requirements (albumin, Ca(2+), HCO(3)(-)).
- Measurement of intracellular cAMP levels and protein tyrosine phosphorylation.
Main Results:
- GalT I-null sperm exhibit precocious capacitation, showing maximal zona pellucida binding and heightened acrosome reaction sensitivity without in vitro capacitation.
- Loss of GalT I impairs binding to decapacitating epididymal glycoconjugates.
- Capacitation in GalT I-null sperm is independent of albumin, Ca(2+), and HCO(3)(-), with elevated cAMP levels.
- Protein tyrosine phosphorylation and motility kinetics remain unaltered in GalT I-null sperm compared to wild-type.
Conclusions:
- GalT I functions as a negative regulator of sperm capacitation, primarily by suppressing intracellular signaling pathways in the sperm head.
- The absence of GalT I leads to constitutive activation of pathways normally promoted by capacitation co-factors.
- These findings offer new insights into the molecular control of sperm maturation and fertilization.
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